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A General And Rapid Cell-Free Approach For The Interrogation Of Protein–Protein, Protein–Dna, And Protein–Rna Interactions And Their Antagonists Utilizing Split-Protein Reporters, Jason R. Porter, Cliff I. Stains, Benjamin W. Jester, Indraneel Ghosh
A General And Rapid Cell-Free Approach For The Interrogation Of Protein–Protein, Protein–Dna, And Protein–Rna Interactions And Their Antagonists Utilizing Split-Protein Reporters, Jason R. Porter, Cliff I. Stains, Benjamin W. Jester, Indraneel Ghosh
Cliff Stains Publications
Split-protein reporters have emerged as a powerful methodology for imaging biomolecular interactions which are of much interest as targets for chemical intervention. Herein we describe a systematic evaluation of split-proteins, specifically the green fluorescent protein, β-lactamase, and several luciferases, for their ability to function as reporters in completely cell-free systems to allow for the extremely rapid and sensitive determination of a wide range of biomolecular interactions without the requirement for laborious transfection, cell culture, or protein purification (12–48 h). We demonstrate that the cell-free split-luciferase system in particular is amenable for directly interrogating protein–protein, protein–DNA, and protein–RNA interactions in homogeneous …